4 research outputs found
Generation and characterization of induced pluripotent stem cell line IITGi001-A derived from adult human primary dermal fibroblasts
Adult human primary dermal fibroblasts [ATCC (PCS-201-012)] were reprogrammed by transfection of oriP/EBNA-1 based episomal plasmids expressing OCT3/4, SOX2, KLF4, L-MYC, LIN28 and a p53 shRNA (Okita et al., 2011) to give rise to induced pluripotent stem cells (iPSCs). These iPSCs expressed core pluripotency markers, maintained normal karyotype, and showed tri-lineage differentiation potential. Further, the absence of episomal plasmid integration in this iPSC line was confirmed by genomic PCR. In addition, DNA fingerprinting of fibroblast and iPSC DNA by microsatellite analysis confirmed the genetic identity of this cell line. This iPSC line was shown to be free from mycoplasma contamination
Population Pharmacokinetics of Cyclophosphamide in Patients with Thalassaemia Major Undergoing HSCT Shows Body Weight, CYP450, GST and ALDH Polymorphisms as Covariates Explaining Inter-Individual Variation.
Identification of novel HPFH-like mutations by CRISPR baseediting that elevate the expression of fetal hemoglobin
Naturally occurring point mutations in the HBG promoter switch
hemoglobin synthesis from defective adult beta-globin to fetal
gamma-globin in sickle cell patients with hereditary persistence
of fetal hemoglobin (HPFH) and ameliorate the clinical severity.
Inspired by this natural phenomenon, we tiled the highly
homologous HBG proximal promoters using adenine and cytosine
base editors that avoid the generation of large deletions and
identified novel regulatory regions including a cluster at the
-123 region. Base editing at -123 and -124 bp of HBG promoter
induced fetal hemoglobin (HbF) to a higher level than disruption
of well-known BCL11A binding site in erythroblasts derived from
human CD34+ hematopoietic stem and progenitor cells (HSPC). We
further demonstrated in vitro that the introduction of -123T > C
and -124T > C HPFH-like mutations drives gamma-globin expression
by creating a de novo binding site for KLF1. Overall, our
findings shed light on so far unknown regulatory elements within
the HBG promoter and identified additional targets for
therapeutic upregulation of fetal hemoglobin.ISSN:2050-084